Phlorotannins from Brown Algae: inhibition of advanced glycation end products formation in high glucose induced Caenorhabditis elegans.

Advanced Glycation End products (AGE) generated in a non enzymatic protein glycation process are frequently associated with diabetes, aging and other chronic diseases. Here, we explored the protective effect of phlorotannins from brown algae Padina pavonica, Sargassum polycystum and Turbinaria ornata against AGEs formation. Phlorotannins were extracted from brown algae with methanol and its purity was analyzed by TLC and RP-HPLC-DAD. Twenty five grams of P. pavonica, S. polycystum, T. ornata yielded 27.6 ± 0.8 µg/ml, 37.7 µg/ml and 37.1 ± 0.74 µg/ml of phloroglucinol equivalent of phlorotannins, respectively. Antioxidant potentials were examined through DPPH assay and their IC50 values were P. pavonica (30.12 ± 0.99 µg), S. polycystum (40.9 ± 1.2 µg) and T. ornata (22.9 ± 1.3 µg), which was comparatively lesser than the control ascorbic acid (46 ± 0.2 µg). Further, anti-AGE activity was examined in vitro by BSA-glucose assay with the extracted phlorotannins of brown algae (P. pavonica, 15.16 ± 0.26 µg/ml; S. polycystum, 35.245 ± 2.3 µg/ml; T. ornata, 22.7 ± 0.3 µg/ml), which revealed the required concentration to inhibit 50% of albumin glycation (IC50) were lower for extracts than controls (phloroglucinol, 222.33 ± 4.9 µg/ml; thiamine, 263 µg/ml). Furthermore, brown algal extracts containing phlorotannins (100 µl) exhibited protective effects against AGE formation in vivo in C. elegans with induced hyperglycemia.

Correction to "Nitrogen-Doped Graphene Quantum-Dot Combined Sodium 10-Amino-2-methoxyundecanoate: Studies of Proinflammatory Gene Expression and Live Cell Imaging".

[This corrects the article DOI: 10.1021/acsomega.8b02085.].

Nitrogen-Doped Graphene Quantum Dot-Combined Sodium 10-Amino-2-methoxyundecanoate: Studies of Proinflammatory Gene Expression and Live Cell Imaging.

Marine cyanobacteria are renowned for producing bioactive secondary metabolites with great structural diversity via mixed biosynthetic pathways. Lyngbya sp., a marine cyanobacterium, produces many metabolites with anti-inflammatory potentials; nevertheless, its bioactive metabolites exercising providing protection against inflammation has been deciphered inadequate. In this study, the ethanolic fraction of the Lyngbya sp. extract was purified and identified as sodium 10-amino-2-methoxyundecanoate (SAM) using Fourier-transform infrared spectroscopy, nuclear magnetic resonance, and electron spray ionization-mass spectroscopy. SAM showed prominent inhibition of inflammation, which was analyzed by reactive oxygen species generation and nitric oxide (NO) inhibition assay. Furthermore, the anti-inflammatory potentials of SAM were evaluated in lipopolysaccharide (LPS)-induced RAW 264.7 macrophage cell lines by fluorescence-activated cell sorting analysis, which evidenced prominent decrease in COX-2 expression (∼90%) with SAM-treated cells than the control. Subsequently, a semiquantitative real-time polymerase chain reaction analysis also revealed the downregulation of COX-2, iNOS, TNF-α, NF-κß, IL-1α, IL-1ß, IL-4, and IL-6 gene expression in SAM-treated LPS-induced RAW 264.7 cells. To further enhance the delivery of SAM into the cells, it was combined with N-doped graphene quantum dots (N-GQDs) for the anti-inflammatory potentials. It resulted in improved downregulation of COX-2, iNOS, TNF-α, NF-κß, IL-1α, IL-1ß, IL-4, and IL-6 than cells treated with SAM alone. Conclusively, N-GQDs combined with SAM have the effective therapeutic potential as an inhibitor of inflammation by modulating the expression of different cytokine genes.

Cloning and expression of L-asparaginase from Bacillus tequilensis PV9W and therapeutic efficacy of Solid Lipid Particle formulations against cancer.

L-asparaginase, a therapeutic involved in cancer therapy, from Bacillus tequilensis PV9W (ansA gene) was cloned and over expressed in Escherichia coli BL21 (DE3), achieved the aim of maximizing the yield of the recombinant enzyme (6.02 ± 1.77 IU/mL) within 12 h. The native L-asparaginase of B. tequilensis PV9W was encapsulated using solid lipid particles by hot lipid emulsion method, which is reported for first time in this study. Subsequently, the lipid encapsulated L-asparaginase (LPE) was characterized by SEM, UV-Vis spectroscopy, FT-IR, SDS-PAGE and its thermo stability was also analyzed by TGA. Further characterization of LPE revealed that enzyme was highly stable for 25 days when stored at 25 °C, showed high pH (9) tolerance and longer trypsin half-life (120 min). In addition, the cytotoxic ability of LPE on HeLa cells was highly enhanced compared to the native L-asparaginase from Bacillus tequilensis PV9W. Moreover, better kinetic velocity and lower Km values of LPE aided to detect L-asparagine in cell extracts by Differential Pulse Voltammetry (DPV) method. The LPE preparation also showed least immunogenic reaction when tested on normal macrophage cell lines. This LPE preparation might thus pave way for efficient drug delivery and enhancing the stability of L-asparaginase for its therapeutic applications.

Agro Waste Utilization for Cost-Effective Production of l-Asparaginase by Pseudomonas plecoglossicida RS1 with Anticancer and Acrylamide Mitigation Potential.

Agricultural wastes such as the peels of onion and garlic were used as a supplement along with l-asparagine for the very first time to produce increased yield of l-asparaginase by Pseudomonas plecoglossicida RS1. Statistical optimization strategies such as response surface methodology were used to generate a medium composition containing extracts of 0.9 (v/v) of garlic peel waste and 0.5% (v/v) onion peel waste along with 0.2% (w/w) l-asparagine, which yielded a twofold increase in the enzyme activity compared to the unsupplemented minimal (M-9) medium. The presence of l-asparagine content in the peel extract was confirmed by high-performance liquid chromatography. Further, l-asparaginase was purified to homogeneity, and identity was confirmed by matrix-assisted laser desorption ionization time-of-flight analysis. The application of the purified l-asparaginase as a therapeutic was studied in HeLa cells which showed a p53-mediated G2 cell cycle arrest. Moreover, the purified l-asparaginase showed effective acrylamide mitigation in vitro, at 6 IU, and its effective degradation was also demonstrated by the effect on chemotactic index of Caenorhabditis elegans and the restoration of the cognitive abilities of C. elegans which was coexposed to acrylamide and l-asparaginase compared to that exposed to acrylamide alone. Thus, l-asparaginase, with multipotent applications, was produced by effective waste utilization for economical commercial production.

Phlorotannins from Brown Algae: inhibition of advanced glycation end products formation in high glucose induced Caenorhabditis elegans.

Advanced Glycation End products (AGE) generated in a non enzymatic protein glycation process are frequently associated with diabetes, aging and other chronic diseases. Here, we explored the protective effect of phlorotannins from brown algae Padina pavonica, Sargassum polycystum and Turbinaria ornata against AGEs formation. Phlorotannins were extracted from brown algae with methanol and its purity was analyzed by TLC and RP-HPLC-DAD. Twenty five grams of P. pavonica, S. polycystum, T. ornata yielded 27.6±0.8 µg/ml, 37.7 µg/ml and 37.1±0.74 µg/ml of phloroglucinol equivalent of phlorotannins, respectively. Antioxidant potentials were examined through DPPH assay and their IC50 values were P. pavonica (30.12±0.99 µg), S. polycystum (40.9±1.2 µg) and T. ornata (22.9±1.3 µg), which was comparatively lesser than the control ascorbic acid (46±0.2 µg). Further, anti-AGE activity was examined in vitro by BSA-glucose assay with the extracted phlorotannins of brown algae (P. pavonica, 15.16±0.26 µg/ml; S. polycystum, 35.245±2.3 µg/ml; T. ornata, 22.7±0.3 µg/ml), which revealed the required concentration to inhibit 50% of albumin glycation (IC50) were lower for extracts than controls (phloroglucinol, 222.33±4.9 µg/ml; thiamine, 263 µg/ml). Furthermore, brown algal extracts containing phlorotannins (100 µl) exhibited protective effects against AGE formation in vivo in C. elegans with induced hyperglycemia.